Cationic amphiphilic drugs cause a marked expansion of apparent lysosomal volume: implications for an intracellular distribution-based drug interaction.
Cationic amphiphilic drugs cause a marked expansion of apparent lysosomal volume: implications for an intracellular distribution-based drug interaction.
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DOI：
10.1021/mp200641e
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发表时间：
2012-05-07
影响因子：
4.9
通讯作者：
Krise JP
中科院分区：
文献类型：
--
作者：
Funk RS;Krise JP
药物在高度分隔的哺乳动物细胞内的分布方式会影响其活性和药代动力学行为。许多市售药物被认为是亲溶酶体的，这意味着它们通过离子捕获型机制大量积聚在溶酶体中。亲溶酶体药物通常具有非常大的表观分布容积和体内较长的半衰期，尽管与脂肪组织的结合极少。在本报告中，我们检验了一种药物（作用药物）在溶酶体中的积聚可能影响后续给予的另一种药物（受影响药物）的积聚，从而导致基于细胞内分布的药物相互作用这一预测。为了验证这一假设，细胞在10μM浓度下暴露于9种不同的含疏水胺的药物，包括丙咪嗪、氯丙嗪和胺碘酮，持续24至48小时。在暴露于作用药物后，对溶酶体示踪红（LTR）这种亲溶酶体模型探针的细胞积聚情况进行了定量和显微镜评估。我们发现所有测试的作用药物都导致LTR的细胞积聚显著增加。细胞暴露于丙咪嗪导致其他亲溶酶体探针和药物的细胞积聚增加，包括溶酶体示踪绿、柔红霉素、普萘洛尔和甲胺；然而，丙咪嗪没有改变非亲溶酶体含胺分子的细胞积聚，包括线粒体示踪红和磺酰罗丹明101。在使用离子载体消除细胞内pH梯度的研究中，我们能够区分基于离子捕获的细胞积聚和与pH梯度无关的残留积聚。这些评估结果结合溶酶体pH测量使我们能够估计丙咪嗪处理前后细胞溶酶体的相对水体积。我们的结果表明，丙咪嗪暴露导致溶酶体体积扩大4倍，这为观察到的药物相互作用提供了基础。丙咪嗪诱导的溶酶体体积扩大被证明是时间和温度依赖性的，并且通过使细胞暴露于羟丙基 - β - 环糊精可逆转，这降低了溶酶体的胆固醇负荷。这表明溶酶体体积的扩大是在作用药物诱导的溶酶体胆固醇含量升高之后发生的。为支持这一说法，从尼曼 - 匹克C型病患者分离的细胞中LTR的细胞积聚被证明更高，已知这些细胞在溶酶体中胆固醇过度积聚。
How a drug distributes within highly compartmentalized mammalian cells can affect both the activity and pharmacokinetic behavior. Many commercially available drugs are considered to be lysosomotropic, meaning they are extensively sequestered in lysosomes by an ion trapping-type mechanism. Lysosomotropic drugs typically have a very large apparent volume of distribution and a prolonged half-life in vivo, despite minimal association with adipose tissue. In this report we tested the prediction that the accumulation of one drug (perpetrator) in lysosomes could influence the accumulation of a secondarily administered one (victim), resulting in an intracellular distribution-based drug interaction. To test this hypothesis cells were exposed to nine different hydrophobic amine-containing drugs, which included imipramine, chlorpromazine and amiodarone, at a 10 µM concentration for 24 to 48 hours. After exposure to the perpetrators the cellular accumulation of LysoTracker Red (LTR), a model lysosomotropic probe, was evaluated both quantitatively and microscopically. We found that all of the tested perpetrators caused a significant increase in the cellular accumulation of LTR. Exposure of cells to imipramine caused an increase in the cellular accumulation of other lysosomotropic probes and drugs including LyosTracker Green, daunorubicin, propranolol and methylamine; however, imipramine did not alter the cellular accumulation of non-lysosomotropic amine-containing molecules including MitoTracker Red and sulforhodamine 101. In studies using ionophores to abolish intracellular pH gradients we were able to resolve ion trapping-based cellular accumulation from residual pH-gradient independent accumulation. Results from these evaluations in conjunction with lysosomal pH measurements enabled us to estimate the relative aqueous volume of lysosomes of cells before and after imipramine treatment. Our results suggest that imipramine exposure caused a 4-fold expansion in the lysosomal volume, which provides the basis for the observed drug interaction. The imipramine-induced lysosomal volume expansion was shown to be both time- and temperature-dependent and reversed by exposing cells to hydroxypropyl-β-cyclodextrin, which reduced lysosomal cholesterol burden. This suggests that the expansion of lysosomal volume occurs secondary to perpetrator-induced elevations in lysosomal cholesterol content. In support of this claim, the cellular accumulation of LTR was shown to be higher in cells isolated from patients with Niemann-Pick Type C disease, which are known to hyper-accumulate cholesterol in lysosomes.
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影响因子：
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影响因子：
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